Spectrum-Effect Relationships between High-Performance Liquid Chromatography (HPLC) Fingerprints and the Antioxidant and Anti-Inflammatory Activities of Collagen Peptides.

Junwen Wang,Wei Liu,Xiangliang Yang,Ming Liang,Ting Zhang,Dan Luo,Xiquan Yin,Ying Zhang

doi:10.3390/molecules23123257

Abstract

A total of 13 batches of collagen peptide samples were extracted, isolated, and purified from chicken sternal cartilage under various process parameters. The fingerprint profiles of 13 batches of collagen peptides were established by high-performance liquid chromatography (HPLC). In addition, the amino acid profiles and molecular weight distributions of collagen peptides were investigated. The in vitro antioxidant activities of the peptide samples were measured using the 2,2′-Azinobis (3-ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) assay, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, the ferric-reducing antioxidant power (FRAP) assay and an assay of the oxidative damage induced by hydrogen peroxide (H2O2) in the degenerative cartilage cells from the knee joint of rat C518 (C518 cell line). The anti-inflammatory activities of the peptide samples were assessed by measuring the inflammatory responses induced by lipopolysaccharides (LPS) in C518 cells. Subsequently, the spectrum-effect relationships between HPLC fingerprints and the antioxidant and anti-inflammatory activities of collagen peptides were investigated using grey relational analysis (GRA). Fifteen common peaks were obtained from the HPLC fingerprints of collagen peptides. Each collagen peptide sample had a characteristic set of amino acid types and contents. All of the hydrolysates of the collagen peptides were primarily composed of fractions II (500–1000 Da) and III (1000–3000 Da). Collagen peptides exhibited good scavenging activity on ABTS radical, DPPH radical, and ferric-reducing antioxidant power. Collagen peptides were also effective against H2O2-induced cellular oxidative damage in C518 cells. The antioxidant activity of collagen peptides was due to the low molecular weight and the presence of antioxidant and hydrophobic amino acid residues within its sequence. Collagen peptides significantly inhibited the secretion of inflammatory cytokines IL-1β, TNF-α, and PGE2 in C518 cells. The anti-inflammatory activity of collagen peptides may include increased synthesis of the key components of extracellular matrix (ECM) and inhibited apoptosis of chondrocytes. The GRA results showed that peaks 2, 3, and 8 were the main components contributing to the antioxidant activity of the collagen peptides, whereas peaks 11 and 14 were the main components contributing to the anti-inflammatory activity of the collagen peptides. The components of peaks 8 and 14 were identified as GPRGPPGPVGP and VAIQAVLSLYASGR by UPLC-MS/MS. Those identified collagen peptides offer a potential therapeutic strategy for the treatment of osteoarthritis (OA) due to their antioxidative stress and due to them disturbing the catabolism and anabolism processes in arthrodial cartilage.

Highlights

Osteoarthritis (OA), the most common degenerative joint disease worldwide found in elderly individuals, is a leading cause of physical disability
The immunologic response to orally administered antigens occurs in the gut-associated lymphoid tissue (GALT), which is composed of epithelium, lamina propria, Peyer’s patch (PP), and mesenteric lymph nodes (MLN)
Peak 7 (Arg, t = 13.881 min) was selected as the reference peak because it was a medium peak in the middle of the chromatogram

Summary

Introduction

Osteoarthritis (OA), the most common degenerative joint disease worldwide found in elderly individuals, is a leading cause of physical disability. Taken native CII antigens interact with PP in the GALT, resulting in turning off the T-cell attack to CII in the cartilage. This desensitization process in PP, known as oral tolerance, avoids the recognition of endogenous CII in the cartilage as an antigen by the immune system [2]. The effects of oral administration of CII obtained from chicken, bovine, and sheep sources have been evaluated for the treatment of arthritis [3]. Chicken sternal cartilage containing a high amount of collagen is recognized as a potential source of CII [4]. Derived CII from chick sternal cartilage is thought to be a potential oral alternative for OA treatment

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